Is it possible that your getting energy transfer?
 
________________________________________________________________________________
 
 
Paul Goodwin
Image Analysis Lab
FHCRC, Seattle, WA
 
On Thu, 23 Feb 1995, Craig Daly wrote:
 
> Hi Folks,
>
> I have a couple of results which I am having trouble explaining and I hope
> someone can put me right.  I use live tissue (blood vessels and smooth
> muscle cells) on our confocal and recently have been using the probes
> Hoechst 33342 (UV,nuclei), FITC-BSA (extracellular) and BODIPY FL-prazosin
> (an alpha1-adrenoceptor ligand).  When I use Hoechst with either FITC or
> BODIPY I often observe nuclear staining in the 488/515 range.  From the
> mole probes handbook it would appear that the FITC and BODIPY may be
> binding to thiolated nucleic acid.  My questions are;
>
> i.      Why do I only see this when I have double labelled with Hoechst and
> a                   Fluorescein.
> ii.     How does nucleic acid become thiolated.
> iii.    How does BODIPY and FITC get into live cells (receptor
> internalisation or membrane transport?) both probes are supposed to be
> impermeant.
>
> If you are still reading this, do you use Fluorescent ligands for
> receptors?  If so are you interested in receptor localisation and
> pharmacology?  We are trying to develop a method of real time ligand
> binding using fluorescent probes for receptors and would like to make
> contact with anyone who has a similar interest.
>
> Thanks for taking the time to read this.
>
> Craig.
>
>
>
> ------------------------------------------------------------
> Craig J. Daly
> Research Associate,
> CRI Heart Failure,
> Institute of Physiology,
> University of Glasgow,
> Glasgow G12 8QQ                 Tel. 44 41 339 8855 ext 6606
>                                 Fax. 44 41 330 4100
>                                 [log in to unmask]
> ------------------------------------------------------------
>